树轮生态学研究中微树芯石蜡切片制作的方法探讨

doi:10.3724/SP.J.1258.2013.00100

摘要/Abstract

摘要：

树轮气候学及树轮生态学研究在国内外取得了长足进展, 但对树轮与气候的响应机制缺乏深入的了解, 迫切需要进行以微树芯石蜡切片为基础的树轮生态学研究。形成层活动监测从生理生态学角度出发, 广泛用于树轮对气候变化响应的机制探讨研究中, 然而目前还没有对微树芯石蜡切片制作的方法作详细的论述和探讨。该文通过对祁连圆柏(Sabina przewalskii)等5个不同树种微树芯石蜡切片的制作实践, 对已有的植物石蜡切片技术进行改进, 增加组织软化的步骤, 并对其他步骤作了相应的调整, 详细介绍了微树芯石蜡切片制作的方法, 为树轮生态学、解剖学, 以及木材解剖学关于木质化材料石蜡切片的制作方法提供参考。

关键词: 树轮生态学, 微树芯, 石蜡切片, 树木年轮

Abstract:

Aims Despite numerous dendroclimatological and dendroecological studies conducted over many regions around the world, associated research on the mechanism of how tree species respond to climate change rarely have been reported. In order to explore the climate-growth patterns in detail, observations of cambial activities have been widely used for physiological and ecological insights.
Methods By investigating seasonal dynamics of cambial activities of five different tree species like Sabina przewalskii, Betula albo-sinensis, Pinus tabuliformis, Picea crassifolia, and Picea wilsonii, we improved the pre-existing technique using plant paraffin sections.
Important findings We considered the process of softening and the timing control of dehydration, cleaning, wax-dipping and staining to improve cambial slices and thereby a provide reliable foundation of paraffin sections of lignified samples useful to the domains of dendroclimatology, dendroecology and wood anatomy.

Key words: dendroecology, micro-core, paraffin section, tree-ring

张军周, 勾晓华, 赵志千, 刘文火, 张芬, 曹宗英, 周非飞. 树轮生态学研究中微树芯石蜡切片制作的方法探讨. 植物生态学报, 2013, 37(10): 972-977. DOI: 10.3724/SP.J.1258.2013.00100

ZHANG Jun-Zhou, GOU Xiao-Hua, ZHAO Zhi-Qian, LIU Wen-Huo, ZHANG Fen, CAO Zong-Ying, ZHOU Fei-Fei. Improved method of obtaining micro-core paraffin sections in dendroecological research. Chinese Journal of Plant Ecology, 2013, 37(10): 972-977. DOI: 10.3724/SP.J.1258.2013.00100

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链接本文: https://www.plant-ecology.com/CN/10.3724/SP.J.1258.2013.00100

https://www.plant-ecology.com/CN/Y2013/V37/I10/972

图/表 6

图1 微树芯采样。A, 祁连山东部祁连圆柏采样点。B, 为避免对下次采样产生影响, 采用“Z”字形方式采样。C, 采集的微树芯。D, 微树芯采集后迅速固定。

Fig. 1 Micro-core sampling. A, Sampling points of Sabina przewalskii in eastern Qilian Mountains. B, In order not to affect the next sampling, using “Z” font sampling. C, Micro-core collection. D, Micro-core rapid fixation after collection.

表1 微树芯脱水、透明、浸蜡的步骤及时间

Table 1 Steps and time of micro-cores dehydration, cleaning and waxdip

步骤 Step	试剂 Reagent	时间 Time
1	30%乙醇 30% ethanol	90-120 min
2	50%乙醇 50% ethanol	90-120 min
3	70%乙醇 70% ethanol	90-120 min
4	85%乙醇 85% ethanol	90-120 min
5	95%乙醇 95% ethanol	60-90 min
6	无水乙醇I Anhydrous ethanol I	60-90 min
7	无水乙醇II Anhydrous ethanol II	60 min
8	二甲苯:乙醇= 1:1 Dimethylbenzene: ethanol = 1:1	60 min
9	二甲苯I Dimethylbenzene I	60 min
10	二甲苯II Dimethylbenzene II	60 min
11	二甲苯:石蜡= 1:1 Dimethylbenzene: paraffin = 1:1	48-72 h
12	石蜡I Paraffin I	120 min
13	石蜡II Paraffin II	60 min
14	石蜡III Paraffin III	30 min

表2 微树芯石蜡切片番红-固绿染色步骤及时间

Table 2 Steps and time of micro-cores staining with safranine and fast green

步骤 Step	试剂 Reagent	染色时间 Staining time
1	二甲苯 Dimethylbenzene	12 min
2	无水乙醇:二甲苯= 1:1 Anhydrous ethanol:dimethylbenzene = 1:1	6 min
3	无水乙醇 Anhydrous ethanol	6 min
4	95%乙醇 95% ethanol	6 min
5	85%乙醇 85% ethanol	6 min
6	70%乙醇 70% ethanol	6 min
7	50%乙醇 50% ethanol	6 min
8	30%乙醇 30% ethanol	6 min
9	水 Water	6 min
10	1%番红水溶液 1% safranine water solution	3-5 h
11	水 Water	60 s
12	30%乙醇 30% ethanol	60 s
13	50%乙醇 50% ethanol	60 s
14	70%乙醇 70% ethanol	60 s
15	85%乙醇 85% ethanol	60 s
16	0.5%固绿乙醇溶液 0.5% fast green ethanol solution	30 s
17	95%乙醇 95% ethanol	60 s
18	无水乙醇 Anhydrous ethanol	60 s
19	无水乙醇:二甲苯=1:1 Anhydrous ethanol:dimethylbenzene = 1:1	60 s
20	二甲苯 Dimethylbenzene	60 s

图2 微树芯切片(A)粘片后(B)和经番红-固绿复染后的切片(C)。

Fig. 2 Paraffin sections of micro-cores (A), bonding (B) and slides of stained with safranin and fast green (C).

图3 番红-固绿染色后的祁连圆柏微树芯显微图片。C, 形成层区域; L, 木质化的细胞; nX, 当年新形成的木质部; P, 韧皮部; X, 上一年的木质部。

Fig. 3 Micro-cores micrograph of Sabina przewalskii stained with safranin-fast green. C, cambial zone; L, lignified cells; nX, current year newly formed xylem; P, phloem; X, last year xylem.

图4 不同树种微树芯石蜡切片显微图片。A, 红桦。B, 油松。C, 青海云杉。D, 青杄。标尺=100 μm。

Fig. 4 Micro-cores paraffin section micrograph of different tree species. A, Betula albo-sinensis. B, Pinus tabuliformis. C, Picea crassifolia. D, Picea wilsonii. Bar = 100 μm.

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